Skip to main content
PMC home page
The EMBO Journal logoLink to The EMBO Journal
. 1991 Dec;10(13):4105–4112. doi: 10.1002/j.1460-2075.1991.tb04987.x

The amino-terminal helix of GM-CSF and IL-5 governs high affinity binding to their receptors.

A B Shanafelt 1, A Miyajima 1, T Kitamura 1, R A Kastelein 1
PMCID: PMC453160  PMID: 1836758

Abstract

Transduction of the biological effects of granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-5 (IL-5) requires the interaction of each cytokine with at least two cell surface receptor components, one of which is shared between these two cytokines. A strategy is presented that allowed us to identify receptor binding determinants in GM-CSF and IL-5. Mixed species (human and mouse) receptors were used to locate unique receptor binding domains on a series of human-mouse hybrid GM-CSF and IL-5 cytokines. Results show that the interaction of these two cytokines with the shared subunit of their high affinity receptor complexes is governed by a very small part of their peptide chains. The presence of a few key residues in the amino-terminal alpha-helix of each ligand is sufficient to confer specificity to the interaction. Comparison with other cytokines suggests that the amino-terminal helix of many of these proteins may contain the recognition element for the formation of high affinity binding sites with the alpha subunit of their multi-component receptors.

Full text

PDF
4106

Images in this article

4106Image
on p.4106
4107Image
on p.4107

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Altmann S. W., Johnson G. D., Prystowsky M. B. Single proline substitutions in predicted alpha-helices of murine granulocyte-macrophage colony-stimulating factor result in a loss in bioactivity and altered glycosylation. J Biol Chem. 1991 Mar 15;266(8):5333–5341. [PubMed] [Google Scholar]
  2. Bazan J. F. Haemopoietic receptors and helical cytokines. Immunol Today. 1990 Oct;11(10):350–354. doi: 10.1016/0167-5699(90)90139-z. [DOI] [PubMed] [Google Scholar]
  3. Bazan J. F. Structural design and molecular evolution of a cytokine receptor superfamily. Proc Natl Acad Sci U S A. 1990 Sep;87(18):6934–6938. doi: 10.1073/pnas.87.18.6934. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Brandhuber B. J., Boone T., Kenney W. C., McKay D. B. Three-dimensional structure of interleukin-2. Science. 1987 Dec 18;238(4834):1707–1709. doi: 10.1126/science.3500515. [DOI] [PubMed] [Google Scholar]
  5. Budel L. M., Elbaz O., Hoogerbrugge H., Delwel R., Mahmoud L. A., Löwenberg B., Touw I. P. Common binding structure for granulocyte macrophage colony-stimulating factor and interleukin-3 on human acute myeloid leukemia cells and monocytes. Blood. 1990 Apr 1;75(7):1439–1445. [PubMed] [Google Scholar]
  6. Chiba S., Shibuya K., Piao Y. F., Tojo A., Sasaki N., Matsuki S., Miyagawa K., Miyazono K., Takaku F. Identification and cellular distribution of distinct proteins forming human GM-CSF receptor. Cell Regul. 1990 Mar;1(4):327–335. doi: 10.1091/mbc.1.4.327. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cunningham B. C., Bass S., Fuh G., Wells J. A. Zinc mediation of the binding of human growth hormone to the human prolactin receptor. Science. 1990 Dec 21;250(4988):1709–1712. doi: 10.1126/science.2270485. [DOI] [PubMed] [Google Scholar]
  8. Devos R., Plaetinck G., Van der Heyden J., Cornelis S., Vandekerckhove J., Fiers W., Tavernier J. Molecular basis of a high affinity murine interleukin-5 receptor. EMBO J. 1991 Aug;10(8):2133–2137. doi: 10.1002/j.1460-2075.1991.tb07747.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gearing D. P., King J. A., Gough N. M., Nicola N. A. Expression cloning of a receptor for human granulocyte-macrophage colony-stimulating factor. EMBO J. 1989 Dec 1;8(12):3667–3676. doi: 10.1002/j.1460-2075.1989.tb08541.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gorman D. M., Itoh N., Kitamura T., Schreurs J., Yonehara S., Yahara I., Arai K., Miyajima A. Cloning and expression of a gene encoding an interleukin 3 receptor-like protein: identification of another member of the cytokine receptor gene family. Proc Natl Acad Sci U S A. 1990 Jul;87(14):5459–5463. doi: 10.1073/pnas.87.14.5459. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. HOWARD-FLANDERS P., SIMSON E., THERIOT L. A LOCUS THAT CONTROLS FILAMENT FORMATION AND SENSITIVITY TO RADIATION IN ESCHERICHIA COLI K-12. Genetics. 1964 Feb;49:237–246. doi: 10.1093/genetics/49.2.237. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hayashida K., Kitamura T., Gorman D. M., Arai K., Yokota T., Miyajima A. Molecular cloning of a second subunit of the receptor for human granulocyte-macrophage colony-stimulating factor (GM-CSF): reconstitution of a high-affinity GM-CSF receptor. Proc Natl Acad Sci U S A. 1990 Dec;87(24):9655–9659. doi: 10.1073/pnas.87.24.9655. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Itoh N., Yonehara S., Schreurs J., Gorman D. M., Maruyama K., Ishii A., Yahara I., Arai K., Miyajima A. Cloning of an interleukin-3 receptor gene: a member of a distinct receptor gene family. Science. 1990 Jan 19;247(4940):324–327. doi: 10.1126/science.2404337. [DOI] [PubMed] [Google Scholar]
  14. Kitamura T., Hayashida K., Sakamaki K., Yokota T., Arai K., Miyajima A. Reconstitution of functional receptors for human granulocyte/macrophage colony-stimulating factor (GM-CSF): evidence that the protein encoded by the AIC2B cDNA is a subunit of the murine GM-CSF receptor. Proc Natl Acad Sci U S A. 1991 Jun 15;88(12):5082–5086. doi: 10.1073/pnas.88.12.5082. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kitamura T., Sato N., Arai K., Miyajima A. Expression cloning of the human IL-3 receptor cDNA reveals a shared beta subunit for the human IL-3 and GM-CSF receptors. Cell. 1991 Sep 20;66(6):1165–1174. doi: 10.1016/0092-8674(91)90039-2. [DOI] [PubMed] [Google Scholar]
  16. Kunkel T. A., Roberts J. D., Zakour R. A. Rapid and efficient site-specific mutagenesis without phenotypic selection. Methods Enzymol. 1987;154:367–382. doi: 10.1016/0076-6879(87)54085-x. [DOI] [PubMed] [Google Scholar]
  17. Lopez A. F., Eglinton J. M., Lyons A. B., Tapley P. M., To L. B., Park L. S., Clark S. C., Vadas M. A. Human interleukin-3 inhibits the binding of granulocyte-macrophage colony-stimulating factor and interleukin-5 to basophils and strongly enhances their functional activity. J Cell Physiol. 1990 Oct;145(1):69–77. doi: 10.1002/jcp.1041450111. [DOI] [PubMed] [Google Scholar]
  18. McKenzie A. N., Barry S. C., Strath M., Sanderson C. J. Structure-function analysis of interleukin-5 utilizing mouse/human chimeric molecules. EMBO J. 1991 May;10(5):1193–1199. doi: 10.1002/j.1460-2075.1991.tb08060.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Messing J. New M13 vectors for cloning. Methods Enzymol. 1983;101:20–78. doi: 10.1016/0076-6879(83)01005-8. [DOI] [PubMed] [Google Scholar]
  20. Metcalf D. The molecular biology and functions of the granulocyte-macrophage colony-stimulating factors. Blood. 1986 Feb;67(2):257–267. [PubMed] [Google Scholar]
  21. Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods. 1983 Dec 16;65(1-2):55–63. doi: 10.1016/0022-1759(83)90303-4. [DOI] [PubMed] [Google Scholar]
  22. Munson P. J. LIGAND: a computerized analysis of ligand binding data. Methods Enzymol. 1983;92:543–576. doi: 10.1016/0076-6879(83)92044-x. [DOI] [PubMed] [Google Scholar]
  23. Park L. S., Friend D., Price V., Anderson D., Singer J., Prickett K. S., Urdal D. L. Heterogeneity in human interleukin-3 receptors. A subclass that binds human granulocyte/macrophage colony stimulating factor. J Biol Chem. 1989 Apr 5;264(10):5420–5427. [PubMed] [Google Scholar]
  24. Park L. S., Waldron P. E., Friend D., Sassenfeld H. M., Price V., Anderson D., Cosman D., Andrews R. G., Bernstein I. D., Urdal D. L. Interleukin-3, GM-CSF, and G-CSF receptor expression on cell lines and primary leukemia cells: receptor heterogeneity and relationship to growth factor responsiveness. Blood. 1989 Jul;74(1):56–65. [PubMed] [Google Scholar]
  25. Parry D. A., Minasian E., Leach S. J. Conformational homologies among cytokines: interleukins and colony stimulating factors. J Mol Recognit. 1988 Jun;1(3):107–110. doi: 10.1002/jmr.300010302. [DOI] [PubMed] [Google Scholar]
  26. Ringheim G. E., Freimark B. D., Robb R. J. Quantitative characterization of the intrinsic ligand-binding affinity of the interleukin 2 receptor beta chain and its modulation by the alpha chain and a second affinity-modulating element. Lymphokine Cytokine Res. 1991 Jun;10(3):219–224. [PubMed] [Google Scholar]
  27. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Shanafelt A. B., Johnson K. E., Kastelein R. A. Identification of critical amino acid residues in human and mouse granulocyte-macrophage colony-stimulating factor and their involvement in species specificity. J Biol Chem. 1991 Jul 25;266(21):13804–13810. [PubMed] [Google Scholar]
  29. Shanafelt A. B., Kastelein R. A. Identification of critical regions in mouse granulocyte-macrophage colony-stimulating factor by scanning-deletion analysis. Proc Natl Acad Sci U S A. 1989 Jul;86(13):4872–4876. doi: 10.1073/pnas.86.13.4872. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Zurawski S. M., Imler J. L., Zurawski G. Partial agonist/antagonist mouse interleukin-2 proteins indicate that a third component of the receptor complex functions in signal transduction. EMBO J. 1990 Dec;9(12):3899–3905. doi: 10.1002/j.1460-2075.1990.tb07610.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Zurawski S. M., Zurawski G. Mouse interleukin-2 structure-function studies: substitutions in the first alpha-helix can specifically inactivate p70 receptor binding and mutations in the fifth alpha-helix can specifically inactivate p55 receptor binding. EMBO J. 1989 Sep;8(9):2583–2590. doi: 10.1002/j.1460-2075.1989.tb08397.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES